Difference between revisions of "Part:BBa K2387053"
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In order to test the functionality of this split protein under the araC/pBad promoter, the absorbance and the fluorescence spectra of the full length protein and the split protein were measured after induction with 0.002% arabinose (Figure 1). The stability of the shape in both spectra for the full length protein and the split protein show that the split has been successful.     
 
In order to test the functionality of this split protein under the araC/pBad promoter, the absorbance and the fluorescence spectra of the full length protein and the split protein were measured after induction with 0.002% arabinose (Figure 1). The stability of the shape in both spectra for the full length protein and the split protein show that the split has been successful.     
  
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[[file:T--Wageningen_UR--Results_Spectra.jpg|800px|center|thumb|<p align="justify">'''Figure 1: Absorbance and fluorescence spectra of full length mCerulean (left) and split mCerulean (right), measured after induction with 0.002% arabinose.'''</p>]]
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[[file:T--Wageningen_UR--Results_Spectra.JPG|800px|center|thumb|<p align="justify">'''Figure 1: Absorbance and fluorescence spectra of full length mCerulean (left) and split mCerulean (right), measured after induction with 0.002% arabinose.'''</p>]]
  
 
This protein is part of a collection of split proteins developed for BiFC analysis and tested with the CpxR system ([https://parts.igem.org/Part:BBa_K2387032 BBa_K2387032]). These proteins were characterized through multiple experiments using as interacting proteins antiparallel synthetic leucine zippers. The proteins of this collection are the ones found in Table 1.  
 
This protein is part of a collection of split proteins developed for BiFC analysis and tested with the CpxR system ([https://parts.igem.org/Part:BBa_K2387032 BBa_K2387032]). These proteins were characterized through multiple experiments using as interacting proteins antiparallel synthetic leucine zippers. The proteins of this collection are the ones found in Table 1.  

Revision as of 11:50, 31 October 2017


Split mCerulean controlled by inducible araC/pBAD promoter

This a split version of BBa_K2387052. The two halves of mCerulean are fused to leucine zippers to induce the reassembly. Both parts are under the control of the araC/pBad promoter and each part is under the control of a stron RBS. The reassembly process is irreversible. This part can be used as a positive control for experiments using BiFC, as well as a source for the fragments of mCerulean.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Functional Parameters

In order to test the functionality of this split protein under the araC/pBad promoter, the absorbance and the fluorescence spectra of the full length protein and the split protein were measured after induction with 0.002% arabinose (Figure 1). The stability of the shape in both spectra for the full length protein and the split protein show that the split has been successful.

Figure 1: Absorbance and fluorescence spectra of full length mCerulean (left) and split mCerulean (right), measured after induction with 0.002% arabinose.

This protein is part of a collection of split proteins developed for BiFC analysis and tested with the CpxR system (BBa_K2387032). These proteins were characterized through multiple experiments using as interacting proteins antiparallel synthetic leucine zippers. The proteins of this collection are the ones found in Table 1.

Table 1: Split Proteins.
Protein Part Number (Full length) Part Number (Split Protein)
mRFP BBa_K2387054 BBa_K2387055
eYFP BBa_K2387003 BBa_K2387065
mVenus BBa_K2387045 BBa_K2387046
sfGFP BBa_K2387047 BBa_K2387048
mCerulean BBa_K2387052 BBa_K2387053